骨髓增生异常综合征免疫表型的研究

目的：评价免疫表型测定在骨髓增生异常综合征(MDS)诊断及分型中的价值。方法：应用单克隆抗体方法对55例MDS患者进行免疫表型检测。结果：MDS患者髓系抗原表达明显增高,FAB亚型的抗原表达呈现规律性改变,随着RA向RAEB／RAEB-t转化,较早期的髓系抗原(CD33)逐渐增加,较成熟的CD15抗原和T-淋巴细胞抗原逐渐减少;同时在RAEB/RAEB—t阶段CD34^ 细胞数明显增高;较早期的骨髓细胞表面抗原(CD38、HLA-DR)在MDS表达明显增加。结论：免疫表型的检测对MDS更精确的诊断和分型有重要意义。     

骨髓增生异常综合征的免疫表型特征分析

 【摘要】　目的　评价免疫表型在骨髓增生异常综合征（MDS）诊断中的价值。方法　采用流式细胞术对27例MDS患者的骨髓细胞进行免疫表型检测。结果　随着MDS疾病的进展,CD+34细胞比例逐渐升高,分别为：难治性贫血／环形铁粒幼细胞性难治性贫血（RA／RAS）7.43 %,难治性贫血伴原始细胞增多（RAEB）36.81 %,难治性贫血伴原始细胞增多转化型（RAEB-T）56.45 %,3组差异有统计学意义（F＝51.197,P＝0.000）,且各组间差异均有统计学意义（P＜0.05）;髓系抗原CD33、CD13、HLA-DR表达逐渐增高,CD14、CD15抗原表达随着疾病的进展逐渐降低,3组间差异有统计学意义（P＜0.05）;B淋巴细胞表面抗原CD19、CD10的表达随着疾病进展而降低;T淋巴细胞表面抗原CD7表达随着疾病进展而增高,分别为RA／RAS 2.63 %、RAEB 10.79 % 和RAEB-T 11.00 %,3组间差异有统计学意义（F＝10.439,P＝0.001）,其中RA／RAS组与RAEB组、RAEB-T组之间差异有统计学意义（P＝0.000,P＝0.001）。结论　检测MDS患者骨髓细胞的免疫表型有助于MDS的诊断、分型和判断预后,从而为治疗提供依据。     

维吾尔族和汉族骨髓增生异常综合征患者细胞形态学及免疫表型的对比分析

 【摘要】　目的　探讨新疆地区维吾尔族和汉族骨髓增生异常综合征（MDS）患者的细胞形态学和免疫表型特征。方法　对已确诊的67例MDS患者骨髓涂片进行系统观察分型,记录各系病态造血细胞,并进行流式细胞术（FCM）免疫表型检测。结果　67例MDS患者骨髓细胞的粒、红、巨核三系有不同程度的病态造血,依次为粒系[52例（77.6 %）]、巨核系[44例（65.7 %）]、红系[36例（53.7 %）],维吾尔族和汉族患者骨髓三系中出现病态造血表现的比例相近,两组差异无统计学意义（χ2值分别为1.02、0.30、0.02,均P＞0.05）。67例骨髓细胞病态造血改变类型的发生率依次为单圆核巨核细胞 [36例（53.7 %）]、假Pelger核异常粒细胞 [36例（53.7 %）]、红系巨幼样变[33例（49.3 %）]、粒细胞颗粒减少或缺失[27例（40.3 %）]等,维吾尔族和汉族患者发生率相似。67例MDS患者FCM免疫表型检测结果显示,随着MDS的难治性贫血／难治性贫血伴环形铁粒幼细胞向难治性贫血伴原始细胞过多（RAEB）／转化中的RAEB的进展变化,较成熟的CD15表达率逐渐降低,而较早期的CD34、CD117表达率逐渐升高（χ2值分别为6.23、12.06,8.95、7.37,8.95、8.08,均P＜0.05）,维吾尔族和汉族患者差异无统计学意义（χ2值分别为0.715、0.024、0.146,均P＞0.05）;同时维吾尔族患者CD56表达增高,汉族MDS患者HLA-DR增高,两组比较差异有统计学意义（χ2值分别为3.91、3.90,均P＜0.05）。结论　维吾尔族和汉族MDS患者骨髓细胞形态学病态造血改变相同,多数MDS有两系以上的病态造血。维吾尔族和汉族MDS患者免疫表型抗原表达部分不同,免疫表型的检测对MDS的诊断、分型及预后具有重要意义。     

骨髓增生异常综合征的免疫表型特征分析

Objective To evaluate the value of immunophenotype in diagnosis of myelodysplastic syndrome (MDS).Methods The immunophenotype of bone marrow cells in 27 patients with MDS were detected by monoclonal antibody by flow cytometry.Results As the progression of the disease,CD34 positive cells gradually increased:refractory anemia/ring sideroblasts refractory anemia (RA/ AS) 7.43 %,refractory anemia with excess of blasts (RAEB) 36.81%,refractory anemia with excess of blasts transformed (RAEB-T)56.45 %,and the differences were statistically significant (P ＜0.05); the expressions of CD33+,CD13 and HLA-DR increased gradually,the expressions of CD14 and CD15 antigens gradually decreased,the difference of three groups was statistically significant (P ＜0.05),the differences between RA/RAS and RAEB-T,RAEB and RAEB-T were statistically significant (P ＜0.05); the expression of CD19 and CD10 decreased and the expression of CD7 increased (RA/RAS 2.63 %,RAEB 10.79 % and RAEB-T 11.00 %) with the progression of the disease,the difference of three groups was statistically significant (F =10.439,P ＜0.05),the differences between RA/RAS and RAEB,RA/RAS and RAEB-T were statistically significant (P ＜0.05).Conclusion The detection of immunophenotype of bone marrow cella in patients with MDS contributes to the diagnosis,classification and prognosis of MDS.     

骨髓增生异常综合征的免疫表型特征分析

Objective To evaluate the value of immunophenotype in diagnosis of myelodysplastic syndrome (MDS).Methods The immunophenotype of bone marrow cells in 27 patients with MDS were detected by monoclonal antibody by flow cytometry.Results As the progression of the disease,CD34 positive cells gradually increased:refractory anemia/ring sideroblasts refractory anemia (RA/ AS) 7.43 %,refractory anemia with excess of blasts (RAEB) 36.81%,refractory anemia with excess of blasts transformed (RAEB-T)56.45 %,and the differences were statistically significant (P ＜0.05); the expressions of CD33+,CD13 and HLA-DR increased gradually,the expressions of CD14 and CD15 antigens gradually decreased,the difference of three groups was statistically significant (P ＜0.05),the differences between RA/RAS and RAEB-T,RAEB and RAEB-T were statistically significant (P ＜0.05); the expression of CD19 and CD10 decreased and the expression of CD7 increased (RA/RAS 2.63 %,RAEB 10.79 % and RAEB-T 11.00 %) with the progression of the disease,the difference of three groups was statistically significant (F =10.439,P ＜0.05),the differences between RA/RAS and RAEB,RA/RAS and RAEB-T were statistically significant (P ＜0.05).Conclusion The detection of immunophenotype of bone marrow cella in patients with MDS contributes to the diagnosis,classification and prognosis of MDS.     

TGF-β1对人单核细胞来源树突状细胞的影响

目的探讨TGF-β1对人单核细胞来源DCs活性的影响.方法在诱导DCs成熟时分别加入TGF-β1、IL-2+TGF-β1、CD40L+TGF-β1以及TNF-α+TGF-β1,比较不同细胞因子对DCs的细胞表面标志CD50、CD83、CD80、CD1-α、CD86、HLA-DR和CCR7表达的影响;对DCs分泌IL-12水平的作用以及DCs活化自体T细胞能力的变化.结果TGF-β1组DCs表面标志CD83、CD80、CD86、HLA-DR、CCR7均明显低于其它各组(P＜0.05);IL-12的分泌水平低于其它各组(P＜0.05);激发自体T细胞能力亦较弱(P＜0.05);与其余各组比较,在第10天时TNF-α组DCs表面分子表达较成熟,活化T细胞能力亦较强(P＜0.05).结论TGF-β1对DCs的成熟以及免疫活性具有明显的抑制作用,而TNF-α能够有效逆转TGF-β1的作用.     

伴髓系抗原表达的儿童急性淋巴细胞白血病免疫分型特点与疗效分析

目的 分析伴髓系抗原表达的儿童急性淋巴细胞白血病 (ALL)的免疫分型特点与临床疗效预后的关系.方法 根据国际白血病欧洲协作组 (EGIL)标准,采用流式细胞术检测白血病细胞的免疫表型,对98例初诊急性淋巴细胞白血病(ALL)儿童分为My- -ALL和My+ -ALL组,My+ -ALL包括My+ B-ALL和My+ T-ALL组,给予正规治疗,对其预后进行观察.结果 My+-ALL患儿25例发生率为25.5%(25/98),其中My+ B-ALL 18例,占72.0%(18/25),My+ T-ALL 7例,占28.0%(7/25).My+ B-ALL和My+ T-ALL患儿中,髓系表达指标CD13阳性率最高,其次为CD33,而CD117均未表达.My+ B-ALL与My- B-ALL、My+ T-ALL与My- T-ALL患儿CR率相比较均无统计学意义 (P＞ 0.05),但在My+ B-ALL组患儿达缓解所需的时间差异存在统计学意义(P＜ 0.05).结论 免疫表型的研究为儿童急性白血病亚型提供了临床诊断的依据.My+ -ALL的预后没有因髓系抗原的表达而与My--ALL表现出明显差异.     

乳腺癌区域淋巴结中CD4+T、CD3+/HLA-DR+T细胞的对比分析

目的:探讨乳腺癌区域淋巴结在转移和无转移状态下免疫细胞数量的变化,认识区域淋巴结在机体抗肿瘤中的免疫状态.方法:选择女性非特异性导管癌26例,术中共取腋窝淋巴结42枚,按HE染色结果将淋巴结分为转移和无转移两组,采用流式细胞技术,检测CD4+T、CD3+/HLA-DR+T细胞占总淋巴细胞数的比例,并在两组之间进行比较.结果:42枚淋巴结18枚有转移,24枚无转移；CD4+T细胞在淋巴细胞中占的比例在两组间有显著性差异(P ＜0.001),在发生癌转移的淋巴结中所占的比例明显少于无转移淋巴结；CD3 +/HLA-DR+T细胞在两组间也有显著性差异(P＜0.05),即转移组少于无转移组.结论:乳腺癌腋窝淋巴结发生癌转移后,细胞免疫功能发生改变,在机体抗肿瘤免疫中具有免疫辅助和直接杀伤作用的CD4+T细胞的比例下降,同时CD3 +/HLA-DR+T细胞即能有效抗肿瘤的活化,T细胞的比例也发生下降.     

骨髓增生异常综合征患者外周血循环CD34+细胞计数的临床研究

目的 探讨骨髓增生异常综合征（MDS）患者外周血循环CD34+细胞计数在疾病分型及预后中的意义。方法 流式细胞仪测定16例健康者、40例MDS患者外周血循环CD34+细胞占有核细胞的百分比（简称CD34+细胞百分比）和绝对细胞数。依据WHO MDS诊断标准、染色体核型以及国际预后积分系统（IPSS）将MDS患者分别划分为RA／RARS／RCMD 组和RAEB Ⅰ／RAEB Ⅱ组,染色体良好组、中间组、不良组,中危Ⅰ组、中危Ⅱ组、高危组。结果MDS患者外周血循环CD34+细胞百分比、绝对数分别为0.67％和17.24个／μl,健康者分别为0.03％和1.63个／μl,两者比较差异有统计学意义（P＜0.01）。MDS患者中,RA／RARS／RCMD组CD34+细胞百分比及绝对数分别为0.05％和3.50个／μl,RAEBⅠ／RAEBⅡ组分别为3.09％和81.95个／μl（P＜0.01）;染色体良好组CD34+细胞百分比及绝对数分别为0.05％和3.50个／μl,中间组为1.29％和18.23个／μl,不良组为3.09％和133.10个／μl,随着不良核型的出现,CD34+细胞百分比及绝对数依次增高（P＜0.05）;中危Ⅰ组CD34+细胞百分比及绝对数分别为0.05％和3.50个／μl,中危Ⅱ组为1.57％和35.55个／μl,高危组为8.15％和192.05个／μl,随着IPSS分值的递增,CD34+细胞百分比及绝对数逐渐增高（P＜0.01）。结论 MDS患者外周血循环CD34+细胞计数存在异常增高现象,其计数水平的检测有助于MDS的分型和预后判断。     

淋系抗原表达对急性髓系白血病预后的价值

目的:探讨淋系抗原表达在急性髓系白血病（acute myeloid leukemia,AML）预后方面的临床意义。方法:应用流式细胞术检测我院101例初诊AML患者的免疫表型,以CD7-CD19-CD56-AML为对照组,将CD7+AML组、CD19+AML、CD56+AML组与对照组的临床特征、疗效进行分析比较,随访并观察生存曲线的差异。结果:101例AML患者中淋系抗原表达者52例(51.5%),CD7抗原表达29例（28.7%）,CD56抗原表达29例（28.7%）,CD19抗原表达13例（12.8%）;CD7+AML、CD19+AML、CD56+AML患者与对照组间的发病年龄、肝脾大、髓外浸润、白细胞计数、血红蛋白、骨髓原始细胞比例差异无统计学意义（P＞0.05）,CD7+AML组血小板计数偏低（P＜0.05）;CD56+AML组首次完全缓解率（CR）率及总CR率均低于对照组（P＜0.05）;CD7+AML、CD19+AML组首次CR率及总CR率与对照组相比差异均无统计学意义（P＞0.05）;与对照组相比,CD56+AML组无复发生存期（RFS）缩短（P＜0.05）,CD7+AML、CD19+AML组RFS与对照组差异无统计学意义（P＞0.05）。结论:CD56+AML患者常规化疗不敏感,疗效差,CD56抗原可能是AML患者的预后不良因素;CD7、CD19抗原不影响AML患者的预后。     

Comparative multi-color flow cytometric analysis of cell surface antigens in bone marrow hematopoietic progenitors between refractory anemia and aplastic anemia

Refractory anemia (RA) in myelodysplastic syndrome (MDS) without prominent dysplasia closely resemble the mild type of aplastic anemia (AA) in their hematological features. This sometimes makes it difficult to distinguish clearly between the two diseases. Using the multi-color flow cytometric technique, we compared cell surface antigen expression patterns on bone marrow hematopoietic progenitor cells which were isolated as a CD34 positive- CD45 dull positive with low side scatter intensity (CD34(+)CD45(dull+)SSC(low)) population in flow cytogram between RA (n=12) and AA (n=11). The antigens analyzed in CD34(+)CD45(dull+)SSC(low) mononuclear cells were: CD38 and CD71 for cell growth-related antigens, CD 33 and CD13 for myeloid and monocytoid lineage-associated antigens, CD7 and CD19 for lymphoid lineage, and CD14 for a monocytic lineage specific antigen. The percentages of CD34(+)CD45(dull+)SSC(low) cells in bone marrow non-erythroid mononuclear cells, and the expression frequencies of CD38, CD71, CD33 and CD13 antigens in CD34(+)CD45(dull+)SSC(low) progenitors were all significantly decreased in AA compared to normal bone marrows (n=7) (P<0.005). In contrast, in RA bone marrows the percentages of CD34(+)CD45(dull+)SSC(low) cells showed wide distribution and the cell surface antigen expression patterns varied among each case: some cases showed low frequencies of CD38 and CD71 expression as well as AA, whereas the others showed high expression frequency of specific antigen(s) which may reflect the clonal expansion of an abnormal clone in bone marrow. An MDS patient who had progressed from RA to RAEB showed further projecting pattern of expression of CD38 and CD33 in CD34(+)CD45(dull+)SSC(low) population in accordance with the disease progression. These data suggest that analysis of cell surface antigen expression patterns of CD34(+)CD45(dull+)SSC(low) progenitor cells by multi-color flow cytometry appears to be a useful method for qualitative and quantitative assessment of marrow progenitor states in AA and RA, therefore this method could be helpful for early detection of clonal evolution in MDS.     

骨髓增生异常综合征造血细胞bcl-2 VEGF抗原表达及原位凋亡的研究

  目的  研究骨髓增生异常综合征(MDS)bcl-2、VEGF抗原表达及造血细胞原位凋亡之间的关系, 探讨MDS发病机理。  方法  对MDS患者骨髓活检标本光镜下观察骨髓增生程度、各系造血细胞形态异常情况、不成熟前体细胞异常定位(ALJP)等组织病理学改变, 免疫组织化学染色观测bcl-2及VEGF抗原的表达, TUNEL法检测造血细胞原位凋亡。  结果  13例患者骨髓均有不同程度的异常造血, 异常造血主要累及的细胞系列不尽相同, RA、RAEB两组患者异常造血未见明显区别; 有6例患者检出ALIP现象, 其中RA 3例(3/8, 37.50%), RAEB 3例(3/5, 60.00%); VEGF阳性者占9例(9/13, 69.23%), 其中RA 6例(6/8, 75.00%), RAEB 3例(3/5, 60.00%); bcl-2阳性者占7例, 其中RA4例(4/8, 50.00%), RAEB 3例(3/5, 60.00%); TUNEL检测13例患者均为阳性, 检出ALIP现象者, 其造血细胞发生原位调亡明显, 而bcl-2、VEGF与TUNEL结果之间未见明显关联。  结论  MDS患者均存在不同程度的细胞凋亡异常, 而TUNEL法提供了简便、实用、敏感、快速特异的检测MDS细胞原位凋亡的方法结果初步显示MDS患者ALlP现象明显者其细胞凋亡也明显, 提示调控MDS患者细胞凋亡和恶性克隆增殖可能是两种完全不同的机制。      

Biologic characteristics of 164 patients with myelodysplastic syndromes

Rates of proliferation, apoptosis and cytokine expression were measured in bone marrow (BM) biopsies of 164 myelodysplastic syndrome (MDS) patients. There were 107 males and 57 females. Median age was 69 years and 101 had refractory anemia (RA), 17 RA with ringed sideroblasts (RARS), 38 with RA and excess blasts (RAEB) and 8 with RAEB in transformation (RAEB-t). Apoptosis measured by in-situ end labeling (ISEL) was directly related to the number of macrophages (p = 0.028, n = 83). Mean tumor necrosis factor alpha (TNF-alpha) and ISEL positivity were higher in RAEB + RAEB-t patients (p = 0.0554 and p = 0.06 respectively) while hemoglobin was higher for RA + RARS group (p = 0.0472). Patients with high apoptosis had lower white blood cell counts (p = 0.0009), lower percentage of blasts (p = 0.0009) and higher number of macrophages (p = 0.0086). We conclude that measurements of apoptosis, proliferation and cytokine expression provide important biological information which helps to distinguish RA + RARS patients from RAEB + RAEB-t patients, and may be of additive prognostic significance.     

骨髓增生异常综合征患者骨髓单个核细胞CD34、CD117的表达及其临床意义

 目的　探讨骨髓增生异常综合征（MDS）患者骨髓单个核细胞CD34、CD117的表达及临床意义。方法　采用直接免疫荧光标记法标记细胞表面分化抗原,流式细胞术测定,对37例MDS患者的骨髓单个核细胞CD34、CD117表达及临床意义进行分析。依据MDS的WHO分型方案、染色体核型以及国际预后积分系统（IPSS）将MDS患者划分为RA-RARS-RCMD 组、RAEBⅠ-RAEBⅡ组;染色体良好组、染色体不良组;中危Ⅰ组、中危Ⅱ组、高危组。结果　RA-RARS-RCMD组19例中11例CD34、CD117表达阳性,RAEBⅠ-RAEBⅡ组18例患者CD34、CD117表达均阳性;随着疾病恶性程度的增高,CD34、CD117表达明显升高;染色体良好组22例中14例患者CD34、CD117表达阳性,染色体不良组15例患者均表达CD34、CD117;随着异常克隆的出现,CD34、CD117表达升高;中危Ⅰ组17例中9例CD34、CD117表达阳性;中危Ⅱ组11例、高危组9例,所有患者CD34、CD117表达均阳性;随着预后积分的递增,CD34、CD117表达随之升高。结论　CD34、CD117检测有助于MDS 的分型、预后判断。     

小儿急性白血病免疫表型和骨髓细胞DNA含量及其相互关系的分析

目的　了解小儿急性白血病（ＡＬ）的免疫表型、异倍体、细胞周期分布状况及其之间的关系。方法　应用流式细胞仪检测了４２ 例初诊为ＡＬ患儿的免疫表型及骨髓单个核细胞（ＭＮＣ）的ＤＮＡ含量。结果　４例ＡＬ患者同时表达了Ｂ系和髓系的抗原，预后差。ＡＬ患儿骨髓细胞周期的Ｓ期细胞百分率明显低于正常对照。而且，ＡＬＬ组中，亚二倍体患者的Ｓ期细胞百分率均值与超二倍体患者、二倍体患者比较存在显著差异性，ＡＮＬＬ组中，ＣＤ３３ ／ＣＤ１３ ＜ １ 患者的Ｓ期细胞百分率均值与ＣＤ３３ ／ＣＤ１３ ＞ １ 患者比较亦存在显著差异。结论　ＡＬ患儿的免疫表型、倍体关系、细胞周期及其之间的关系，与临床预后密切相关。     

成人急性髓系白血病的免疫学特点及预后

目的：探讨急性髓细胞白血病（AML）的免疫表型特点。方法：使用淋系和髓系单抗,用间接免疫荧光法对70例原发性AML进行免疫表型分析。结果：所有AML患者的细胞至少被1种髓系单抗标记,各髓系抗原的表达率依次为CD33＞CD13＞CD15。所有M3患者CD9为阳性。16／70例（30％）表达CD34抗原、CD34^ AML组在年龄、外周血象及骨髓原始、幼稚细胞比例等方面与CD34^-组相比较无显著差别,但表达CD34抗原的AML常伴有HLA－DR、CD38、CD7等不成熟细胞表面标记的表达,而较成熟的髓系细胞表面标记CD15则不表达。70例AML中有16例表达淋系抗原,CD4^ 例（13．8％,M2为8．8％,M460％）;CD7^ 9例（16．9％,M1为50％,M218％,M5b16．7％）。CD4^ 的AML患者CD34为低表达,CD33表达。结论：CD9^ 、CD34^-、HLA－DR^-及CD13^ 、CD15^ 是典型M3的免疫表型特点。CD34^ AMLgn AML-M1有着密切的关系,且对化疗反应较差,证明CD34^ 的AML是一组分化程度较差的类型;提示CD7^ 和CD4^ 的AML预后差,CD7^ 的AML的一种独特类型。     

Limited numbers of apoptotic cells in fresh paraffin embedded bone marrow samples of patients with myelodysplastic syndrome

In myelodysplasia (MDS) the precise mechanism of ineffective erythropoiesis is not fully elucidated, but it is suggested that apoptosis may contribute to this process. We performed TdT-mediated dUTP-nick end labelling (TUNEL) staining of paraffin embedded bone marrow specimens to assess the amount of apoptotic cells in 21 MDS patients (7 RA, 3 RARS, 5 RAEB, 3 RAEB-T, 3 CMML) and five normal controls. In 10 MDS patients the TUNEL assay was performed in combination with immunostaining for Glycophorin-A (GpA) to determine apoptosis in the maturing erythroid compartment. To assess the proliferation of the bone marrow cells the expression of Ki-67 antigen was used as a marker. The mean apoptotic index (AI) in MDS patients was not increased (2.3 +/- 3.0% in MDS versus 4.8 +/- 1.2% in normal controls (P < 0.05)). Moreover, no significant difference in mean AI was observed in the GpA+ compartment between MDS and normal controls (0.8 +/- 0.2% versus 0.6 +/- 0.1%). In addition the different FAB-classifications and the different International Prognostic Scoring System (IPSS)-risk groups showed no significant differences between the subgroups. The expression of Ki-67, as marker for proliferative activity, in the GpA+ compartment from MDS did not differ significantly from normal controls (84.0 +/- 12.2% versus 79.9 +/- 20.2%). Our findings suggest that the observed increased apoptosis in in vitro culture assays is related to the detachment of the cells from the microenvironment leading to an increased susceptibility to apoptosis.